SHH Signal Protective Effect In The Primary Cultured Cortical Neurons With Oxidative Stress

Objective Explore the protective effects of SHH signal in the primary cultured cortical neuron with oxidative stress.Methods This experiment was performed in the central laboratory of union hospital and the laboratory of department of neurology, Tongji medical college, Huzhong University of Science and Technology.1.Adult SD male rats were suture model of middle cerebral artery for 3 days for ischemic. Real-time PCR to compare gene expression of Shh, Patched and Gli-1 in the infarct side and the contra lateral noninfarction.2.The gray matter of cerebral cortex dissected from postnatal 1 to 3 days SD rats was dissociated for neurons primary culture. These cultured cells were treated with H2O2 miming oxidative stress. Real-time PCR and western-blot determinate the endogenous expression of Shh and Patched, Gli-1, and explored the origin of Shh.3.Treat primary cultured neurons in oxidative stress condition with Shh and Shh signal inhibitant Cyclopamine, and observed the changes of cell vitality with MTT and cell apoptosis through Hochest and flow cytometry. Results 1.Compared with the contra lateral no infarction, Shh, Patched, Gli-1 expression in the infarction lateral obviously elevated, consistency with related research about Shh expression in the ischemic stroke. 2. Oxidative stress could up-regulate endogenous expression of Shh, Patched, and Gli-1, originated from the neuron cells. 3. Treatment of Shh improved cell vitality and inhibited cell apoptosis, and the effect could be partly inhibited by Cyclopamine.Conclusions Ischemic stroke could improved the expression of Shh and reactive Shh signal, Shh could be originated from neurons. Treatment of Shh could improve cell vitality and inhibit cell apoptosis, playing a role of neuroprotective effect.